Mouse Oncostatin M Recombinant

HomeHematopoietins, Recombinant Mouse Cytokines

Mouse Oncostatin M Recombinant

$70.00 – $4,700.00

SKU: RKP53347 Categories: Hematopoietins, Recombinant Mouse Cytokines Tags: oncostatin-m, OSM, P53347

Description

Methods (2)


accession	P53347

Source	Optimized DNA sequence encoding Mouse Oncostatin-M mature chain was expressed in Escherichia Coli
Molecular weight	Native Mouse Oncostatin-M isgenerated by the proteolytic removal of the signal peptide and propeptide, this molecule has a calculated molecular mass of approximately kDa. Recombinant mouse Oncostatin-M (OSM) is a monomeric protein consisting of 181 amino acid residue subunits, and migrates as an approximately 20 kDa protein under non-reducingand reducing conditions in SDS-PAGE.
Purity	>95%, as determined by SDS-PAGE and HPLC
Biological Activity	The ED(50) was determined by the dose-dependent stimulation of the proliferation of MouseT3 cells is ≤0.5 ng/ml, corresponding to a specific activity of ≥2 x10⁷ units/mg.
Protein Sequence	MQTRLLRTLL SLTLSLLILS MALANRGCSN SSSQLLSQLQ NQANLTGNTE SLLEPYIRLQ NLNTPDLRAA CTQHSVAFPS EDTLRQLSKP HFLSTVYTTL DRVLYQLDAL RQKFLKTPAF PKLDSARHNI LGIRNNVFCM ARLLNHSLEI PEPTQTDSGA SRSTTTPDVF NTKIGSCGFL WGYHRFMGSV GRVFREWDDG STRSR RQSPL RARRKGTRRI RVRHKGTRRI RVRRKGTRRI WVRRKGSRKI RPSRSTQSPT TRA
Endotoxin	Endotoxin content was assayed using a LAL gel clot method. Endotoxin level was found to be less than.1 ng/µg(1EU/µg).
Presentation	Mouse OSM (oncostatin-M) was lyophilized from a 0.2 μm filtered solution in.5% glycine,.5% sucrose,.01% Tween80, mM Glutamic acid, pH.5.
Reconstitution	A quick spin of the vial followed by reconstitution in distilled water to a concentration not less than.1 mg/mL. This solution can then be diluted into other buffers
Storage	The lyophilized protein is stable for at least years from date of receipt at -20° C. Upon reconstitution, this cytokine can be stored in working aliquots at° -° C for one month, or at -20° C for six months, with a carrier protein without detectable loss of activity. Avoid repeated freeze/thaw cycles.
Usage	This cytokine product is for research purposes only.It may not be used for therapeutics or diagnostic purposes.

Interactor	O70458 OSMR_MOUSE
Interactor	Q00560 IL6RB_MOUSE
Biological Process	Growth-regulation
Molecular function	Cytokine

Methods


Impact of OSM on gene expression (A, B) and protein secretion (C, D) in stimulated HSFs. Human recombinant OSM (10 ng/ml) was added with or without 10 μg/ml of neutralizing anti-OSM, or isotype-matched non-specific antibody.
Isolation and culture of Nestin-positive cells from Nes-GFP transgenic mice The testes were dissected from 7-day-old Nes-GFP or C57BL/6 mice, and the tunica albuginea was carefully removed from each testis and minced into small pieces. The interstitial cells were then dissociated from the seminiferous tubules by digestion with 1 mg/ml collagenase type IV in medium'>Dulbecco's medium'>modified medium'>Eagle's medium (DMEM)/F12 at 37 °C for 15 min. After adding DMEM containing 5% fetal bovine serum (FBS, Hyclone) to stop the collagenase activity, the samples were centrifuged at 1 500 rpm for 5 min at room temperature; the pellets were washed twice with phosphate-buffered saline (PBS), resuspended in PBS, and filtered through a 45 μm filter, thereby excluding seminiferous tubules from the preparation. The suspension was passed through a 20 μm strainer, which resulted in single cells. GFP-expressing cells that exhibited fluorescence intensities ∼10-fold greater than the autofluorescence of cells from C57BL/6L mice were enriched… The testes were dissected from 7-day-old Nes-GFP or C57BL/6 mice, and the tunica albuginea was carefully removed from each testis and minced into small pieces. The interstitial cells were then dissociated from the seminiferous tubules by digestion with 1 mg/ml collagenase type IV in medium'>Dulbecco's medium'>modified medium'>Eagle's medium (DMEM)/F12 at 37 °C for 15 min. After adding DMEM containing 5% fetal bovine serum (FBS, Hyclone) to stop the collagenase activity, the samples were centrifuged at 1 500 rpm for 5 min at room temperature; the pellets were washed twice with phosphate-buffered saline (PBS), resuspended in PBS, and filtered through a 45 μm filter, thereby excluding seminiferous tubules from the preparation. The suspension was passed through a 20 μm strainer, which resulted in single cells. GFP-expressing cells that exhibited fluorescence intensities ∼10-fold greater than the autofluorescence of cells from C57BL/6L mice were enriched by FACS using an Influx Cell Sorter and were cultured in ultra-low adherent dishes (Corning). The expansion medium adapted for SLC culture was previously published and was used with modifications; this medium consisted of DE/F12 (1:1) supplemented with 1 ndexamethasone , 1 ng/ml LIF , 5 μg/L insulin-transferrin-sodium selenite , 5% chicken embryo extract (US), 0.1 m-mercaptoethanol , 1% non-essential amino acids , 1% N2 and 2% B27 supplements , 20 ng/ml basic fibroblast growth factor , epidermal growth factor , platelet-derived growth factor , and oncostatin . The cultures were kept at 37 °C in a humidified 5% CO₂ water-jacketed incubator. The medium was changed every 3 days. Read more